Berberine, an isoquinolene alkaloid, together with its related analogs protoberberineand palmatine were detected in cell suspension cultures derived from leaf explants of Tinosporacordifolia. Maximum production of biomass (9.9 g dry cell wt/l) was noticed on day 36 whileberberine was highest (5.5 mg/g dry weight) at day 24. Media pH, plant growth regulators andcarbon sources were optimized for biomass and accumulation of berberine in Linsmaier andSkoog’s medium. Out of 8 cell lines selected, only one line accumulated high berberine (13.9mg/g dry wt) in 4-weeks. While cell aggregates of 500 ?m in diameter promoted production ofbiomass (9.6 g dry cell wt/l), larger cell aggregates (above 500 ?m) favoured berberineaccumulation with 3.8 mg/g dry weight of tissue. The yield of berberine in cell suspensions ofTinospora was 5-14-folds higher than that of intact plants.